Adenosine-mediated Neutrophil Regulation by Dapagliflozin Attenuates Renal Ischemia/Reperfusion Injury: Real-time Imaging and Metabolic Insights

Background. Acute kidney injury resulting from ischemia/reperfusion injury (IRI) remains a significant clinical challenge, with limited therapeutic options. This study investigated the renoprotective mechanisms of dapagliflozin, a sodium-glucose cotransporter 2 inhibitor (SGLT2i), in nondiabetic mice, focusing on neutrophil dynamics and adenosine signaling pathways. Methods. Nondiabetic mice were pretreated with dapagliflozin before bilateral renal IRI. Some groups received concurrent treatment with an adenosine A2A receptor antagonist. We used novel multiphoton intravital imaging, complemented by comprehensive molecular and metabolomic analyses, to visualize neutrophil trafficking during early reperfusion. Results. Dapagliflozin significantly attenuated renal dysfunction ( P < 0.01) and histological damage ( P < 0.01). Real-time imaging revealed that dapagliflozin markedly suppressed neutrophil infiltration into the glomeruli and peritubular capillaries during early reperfusion, and this effect was partially reversed by coadministration of an A2A receptor antagonist. Molecular analyses demonstrated reduced matrix metalloproteinase-9 expression and activity, with decreased levels of endothelial adhesion molecules, including intercellular adhesion molecule-1 and vascular cell adhesion molecule-1. Kidney adenosine levels were significantly increased in dapagliflozin-treated mice and were inversely correlated with matrix metalloproteinase-9 activity. Metabolome analysis revealed significant metabolic reprogramming characterized by suppressed glycolysis, enhanced tricarboxylic acid cycle activity, and elevated adenosine pathway components. Conclusions. Dapagliflozin protects against renal IRI through the adenosine-mediated inhibition of neutrophil infiltration and inflammatory activation. This novel mechanism, involving metabolic reprogramming and enhanced adenosine signaling, extends our understanding of the pleiotropic effects of sodium-glucose cotransporter 2 inhibitors beyond glycemic control and suggests potential therapeutic applications for preventing acute kidney injury in high-risk clinical settings, including kidney transplantation.